ENO1 blockade augment ferroptosis susceptibility in TKIs-resistant CML through GPX4 autophagic degradation

The introduction of tyrosine kinase inhibitors (TKIs) has significantly improved the prognosis of chronic myeloid leukemia (CML), however, approximately 20% CML patients developed resistance to TKIs and lead to treatment failure. Enolase 1 (ENO1) is a critical enzyme involved in glycolysis and was found to be closely related to CML carcinogenesis. Our results indicate ENO1 expression was close correlated with drug responses and disease prognosis in CML. The chemo-resistant CML cell K562/G more relied on glycolysis for energy supply than its sensitive counterparts.,Metabolomic analysis revealed that ENOblock, APIII-α4(AP), synergized with multiple TKIs and induce Ferroptosis in K562/G cells. Bioinformatics analysis suggested that GPX4 play more crucial role in sustaining chemo-resistant CML cell survival. Transcriptomic analysis and WB results revealed GPX4 autophagic degradation induced by ENO1 downregulation. AMPK/mTOR signaling pathway activated by ENO1 downregulationplayed partial role in GPX4 degradation. More importantly, the expression of ENO1 was found to be inversely correlated with that of a transmembrane protein TMEM164.TMEM164 interference would restore the GPX4 autophagic degradation and Ferroptosis susceptibility induced by ENO1 downregulation.Single-cell sequence data revealed a co-expression relationship between ENO1 and GPX4, especially in CML patients with poor TKIs responses. Besides, In vivo animal experiments demonstrated that AP could cooperate with TKIs to relieve the tumor burden with tolerable safety. Taken together, this study demonstrated that ENO1 is a crucial biomarker for CML TKIs responses, and ENO1 blockade could augment TKIs sensitivity and promote the Ferroptosis susceptibility in TKIs-resistant cells by ultimately inducing GPX4 autophagic degradation through AMPK/mTOR pathway and ENO1-TMEM164 interaction, which provide a potential novel target for the clinical treatment of CML.

CML; ENO1; GPX4; chemo-resistant; ferroptosis.